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The blood cell karyotyping method was developed to provide information about chromosomal abnormalities. Lymphocyte cells do not normally undergo subsequent cell divisions. In the presence of a mitogen, lymphocytes are stimulated to enter into mitosis by DNA replication. After 48-72 hours, a mitotic inhibitor is added to the culture to stop mitosis in the metaphase stage. After treatment by hypotonic solution, fixation and staining, chromosomes can be microscopically observed and evaluated for abnormalities.

Remove the supernatant, agitate the cellular sediment and add drop-by-drop 5-10ml of fresh, ice-cold fixative made up of 1 part acetic acid to 3 parts methanol. Leave in 4ºC for 10 minutes.

Repeat steps 7 and 8.

Spin at 500xg for 5 minutes.

Re-suspend the cell pellet in a small volume 0.5-1ml of fresh fixative, drop onto a clean slide and allow to air dry.

At this stage, the preparation can be stained with Orecin or Giemsa. Giemsa banding has become the most widely used technique, and the most common method to obtain this staining is to treat slides with Trypsin-EDTA 10X (Cat. No. 15400-054).